U.S. patent application number 11/884543 was filed with the patent office on 2009-02-19 for cosmetic use of at least one natural ac-n-ser-asp-lys-pro-tetrapeptide or one of its analogues as agent for slowing down loss of hair and/or stimulating hair growth.
This patent application is currently assigned to CENTRE NATIONAL DE LA RECHERCHE SCIENTIFQUE (CNRS. Invention is credited to Joanna Elzbieta Bakala, Jerome Bignon, Jean-Yves Lallemand.
Application Number | 20090048182 11/884543 |
Document ID | / |
Family ID | 34979065 |
Filed Date | 2009-02-19 |
United States Patent
Application |
20090048182 |
Kind Code |
A1 |
Bakala; Joanna Elzbieta ; et
al. |
February 19, 2009 |
Cosmetic Use of at Least One Natural
Ac-N-Ser-Asp-Lys-Pro-Tetrapeptide or One of Its Analogues as Agent
for Slowing Down Loss of Hair and/or Stimulating Hair Growth
Abstract
The invention concerns the cosmetic use of at least one compound
of formula (I) wherein: A.sub.1 is the radical corresponding to D-
or L-Ser; A.sub.2 is the radical corresponding to D- or L-Asp or
Glu; A.sub.3 is the radical corresponding to D- or L-Lys, Arg or
Orn; A.sub.4 is the radical corresponding to D- or L-Pro; R.sub.1,
R.sub.2, R.sub.3 are such as defined in the claims, as agent for
controlling hair loss.
Inventors: |
Bakala; Joanna Elzbieta;
(Paris, FR) ; Bignon; Jerome; (Le Val Saint
Germain, FR) ; Lallemand; Jean-Yves; (Palaiseau,
FR) |
Correspondence
Address: |
FOLEY AND LARDNER LLP;SUITE 500
3000 K STREET NW
WASHINGTON
DC
20007
US
|
Assignee: |
CENTRE NATIONAL DE LA RECHERCHE
SCIENTIFQUE (CNRS
|
Family ID: |
34979065 |
Appl. No.: |
11/884543 |
Filed: |
February 16, 2006 |
PCT Filed: |
February 16, 2006 |
PCT NO: |
PCT/EP2006/060029 |
371 Date: |
August 17, 2007 |
Current U.S.
Class: |
514/8.9 |
Current CPC
Class: |
A61P 17/14 20180101;
A61K 8/64 20130101; A61Q 7/00 20130101 |
Class at
Publication: |
514/18 |
International
Class: |
A61K 38/07 20060101
A61K038/07; A61P 17/14 20060101 A61P017/14 |
Foreign Application Data
Date |
Code |
Application Number |
Feb 18, 2005 |
FR |
0501672 |
Claims
1. A method for slowing down hair loss and/or stimulating hair
growth, comprising applying to the scalp a composition comprising
at least one compound of formula (I): ##STR00002## in which A.sub.1
is the radical corresponding to D- or L-Ser, A.sub.2 is the radical
corresponding to D- or L-Asp or Glu, A.sub.3 is the radical
corresponding to D- or L-Lys, Arg or Orn, A.sub.4 is the radical
corresponding to D- or L-Pro, R.sub.1 and R.sub.2 are independently
chosen from among the hydrogen atom, linear or branched substituted
or non-substituted C.sub.1-C.sub.12 alkyl groups, substituted or
non-substituted linear or branched substituted or non-substituted
C.sub.7-C.sub.20 arylalkyl groups, R.sub.4CO-- and R.sub.4COO--
where R.sub.4 is a linear or branched substituted or
non-substituted C.sub.1-C.sub.12 alkyl group, or a substituted or
non-substituted C.sub.7-C.sub.20 arylalkyl, where substitutions
include OH, NH.sub.2 and COOH, X.sub.1 and X.sub.2 are peptide or
pseudopeptide bonds, X.sub.3 is a radical chosen from among --CO
and --CH.sub.2--, and R.sub.3 is a group chosen from among --OH,
--NH.sub.2, linear or branched alcoxy C.sub.1-C.sub.12 or
--NH--X.sub.4--CH.sub.2-Z, where X.sub.4 is a linear or branched
C.sub.1-C.sub.12 hydrocarbon group and Z is a hydrogen atom or
--OH, --CO.sub.2H or --CONH.sub.2 as well as their physiologically
acceptable salts.
2. The method of claim 1, wherein said compound stimulates the
growth and/or differentiation of stem cells and/or
neo-keratinocytes in the epithelial sheath of hair.
3. The method of claim 1, wherein said compound stimulates the
production of collagen IV and/or laminin 5 in the hair
follicle.
4. The method of claim 1, wherein said compound triggers and/or
stimulates hair growth.
5. The method of claim 1, to increase hair density.
6. The method of claim 1, to prevent alopecia.
7. The method of claim 1, wherein the derivative of formula (I)
includes at least one pseudopeptide bond.
8. The method of claim 1, wherein the compound of formula (I) is
chosen from among: CH.sub.3CO-Ser-Asp-Lys-Pro-OH,
CH.sub.3CO-Ser-.psi.-(CH.sub.2NH)-Asp-Lys-Pro-OH
CH.sub.3CO-Ser-Asp-.psi.-(CH.sub.2NH-Lys-Pro-OH
CH.sub.3CO-Ser-Asp-Lys-.psi.-(CH.sub.2N-Pro-OH
CH.sub.3CO-Ser-.psi.-(CH.sub.2NH)-Asp-Lys-Pro-NH.sub.2
H-Ser-.psi.-(CH.sub.2NH)-Asp-Lys-Pro-OH
H-Ser-Asp-.psi.-(CH.sub.2NH)-Lys-Pro-OH
H-Ser-Asp-Lys-.psi.-(CH.sub.2N)-Pro-OH
HOOCCH.sub.2CH.sub.2CO-Ser-.psi.-(CH.sub.2NH)-Asp-Lys-Pro-OH
HOOCCH.sub.2CH.sub.2CO-Ser-Asp-.psi.-CH.sub.2NH-Lys-Pro-OH
HOOCCH.sub.2CH.sub.2CO-Ser-Asp-Lys-.psi.-(CH.sub.2N-Pro-OH
H-Ser-.psi.-(CH.sub.2NH)-Asp-Lys-Pro-NH.sub.2
H-Ser-Asp-.psi.-(CH.sub.2NH)-Lys-Pro-NH.sub.2
H-Ser-Asp-Lys-.psi.-(CH.sub.2N)-Pro-NH.sub.2
HOOCCH.sub.2CH.sub.2CO-Ser-.psi.-(CH.sub.2NH)-Asp-Lys-Pro-NH.sub.2
HOOCCH.sub.2CH.sub.2CO-Ser-Asp-.psi.-(CH.sub.2NH)-Lys-Pro-NH.sub.2
HOOCCH.sub.2CH.sub.2CO-Ser-Asp-Lys-.psi.-CH.sub.2N-Pro-NH.sub.2
CH.sub.3CO-Ser-Asp-Lys-Pro-NH.sub.2 H-Ser-Asp-Lys-Pro-NH.sub.2
CH.sub.3CO-Ser-Asp-Lys-Pro-NHCH.sub.3 H-Ser-Asp-Lys-Pro-NHCH.sub.3
HOOCCH.sub.2CH.sub.2CO-Ser-Asp-Lys-Pro-NHCH.sub.3
HOOCCH.sub.2CH.sub.2CO-Ser-Asp-Lys-Pro-NH.sub.2
9. The method of claim 1, wherein the compound of formula (I) is
represented by the formula: CH.sub.3CO-Ser-Asp-Lys-Pro-OH.
10. The method of claim 1, wherein the compound of formula (I) is
present in the composition in amounts ranging from 0.001% to 10% by
weight.
11. (canceled)
12. A method for treating and/or preventing alopecia, comprising
applying to the scalp a composition comprising at least one
compound of formula (I): ##STR00003## in which A.sub.1 is the
radical corresponding to D- or L-Ser, A.sub.2 is the radical
corresponding to D- or L-Asp or Glu, A.sub.3 is the radical
corresponding to D- or L-Lys, Arg or Orn, A.sub.4 is the radical
corresponding to D- or L-Pro, R.sub.1 and R.sub.2 are independently
chosen from among the hydrogen atom, linear or branched substituted
or non-substituted C.sub.1-C.sub.12 alkyl groups, substituted or
non-substituted linear or branched substituted or non-substituted
C.sub.7-C.sub.20 arylalkyl groups, R.sub.4CO-- and R.sub.4COO--
where R.sub.4 is a linear or branched substituted or
non-substituted C.sub.1-C.sub.12 alkyl group, or a substituted or
non-substituted C.sub.7-C.sub.20 arylalkyl, where substitutions
include OH, NH.sub.2 and COOH, X.sub.1 and X.sub.2 are peptide or
pseudopeptide bonds, X.sub.3 is a radical chosen from among --CO
and --CH.sub.2--, and R.sub.3 is a group chosen from among --OH,
--NH.sub.2, linear or branched alcoxy C.sub.1-C.sub.12 or
--NH--X.sub.4--CH.sub.2-Z, where X.sub.4 is a linear or branched
C.sub.1-C.sub.12 hydrocarbon group and Z is a hydrogen atom or
--OH, --CO.sub.2H or --CONH.sub.2 as well as their physiologically
acceptable salts.
13. A method for increasing hair density, comprising applying to
the scalp a composition comprising at least one compound of formula
(I): ##STR00004## in which A.sub.1 is the radical corresponding to
D- or L-Ser, A.sub.2 is the radical corresponding to D- or L-Asp or
Glu, A.sub.3 is the radical corresponding to D- or L-Lys, Arg or
Orn, A.sub.4 is the radical corresponding to D- or L-Pro, R.sub.1
and R.sub.2 are independently chosen from among the hydrogen atom,
linear or branched substituted or non-substituted C.sub.1-C.sub.12
alkyl groups, substituted or non-substituted linear or branched
substituted or non-substituted C.sub.7-C.sub.20 arylalkyl groups,
R.sub.4CO-- and R.sub.4COO-- where R.sub.4 is a linear or branched
substituted or non-substituted C.sub.1-C.sub.12 alkyl group, or a
substituted or non-substituted C.sub.7-C.sub.20 arylalkyl, where
substitutions include OH, NH.sub.2 and COOH, X.sub.1 and X.sub.2
are peptide or pseudopeptide bonds, X.sub.3 is a radical chosen
from among --CO and --CH.sub.2-, and R.sub.3 is a group chosen from
among --OH, --NH.sub.2, linear or branched alcoxy C.sub.1-C.sub.12
or --NH--X.sub.4--CH.sub.2-Z, where X.sub.4 is a linear or branched
C.sub.1-C.sub.12 hydrocarbon group and Z is a hydrogen atom or
--OH, --CO.sub.2H or --CONH.sub.2 as well as their physiologically
acceptable salts.
Description
[0001] Cosmetic use of at least one natural tetrapeptide
Ac-N-Asp-Lys-Pro or one of its analogues as an agent for slowing
down loss of hair and/or stimulating hair growth.
[0002] This invention concerns the cosmetic use in a composition of
at least one tetrapeptide compound or analogue as an agent for
slowing down loss of hair and/or stimulating hair growth.
[0003] Humans have around 150 000 hairs on their heads. Each hair
consists of a shaft, the free part that emerges at the surface of
the scalp, and a root (bulb) implanted in the hair follicle. Each
hair has a lifespan ranging from 2 to 7 years.
[0004] Hair is produced by a complex organ, the hair follicle,
consisting of a dermal compartment and an epithelial compartment.
Each of these compartments is subdivided into functional
sub-compartments. The follicles act a <<reservoir >> of
stem cells capable of giving rise to all the cell lines needed to
reconstitute the follicles, epidermis and sebaceous glands.
[0005] In humans, hair growth and renewal are determined mainly by
the activity of hair follicle and the surrounding dermo-epidermal
environment. A follicle is either in the growth phase (production
of the hair shaft) in the course of which hair grows in length
(anagenic phase) or at a stage where growth has stopped (telogenic
phase). After the telogenesis phase and as a result of a
neo-morphogenic process, the follicle is regenerated from the
reservoir of stem cells and initiates a new anagenic phase. With
normal hair which is constantly renewed, about 85% of follicles are
in the growth phase, 2% in the resting phase and over 10% in the
falling out phase.
[0006] Men and women usually lose 50 to 150 hairs a day.
Nevertheless, these hairs are replaced by new growth as a function
of the hair growth cycle. When hair loss becomes visible, this
means that it is excessive (very much in excess of 150 hairs per
day) or, more commonly, because hairs which fall out are replaced
by finer hairs as is the case with individuals with androgenetic
alopecia. In such cases, hair re-growth becomes thinner and thinner
and rarer from one cycle to the next.
[0007] There are many causes of hair loss: diseases such as cancer
or lupus, hormonal changes, stress, certain medications or food
deficiencies. However, in the vast majority of cases, hair loss is
of hereditary or hormonal origin. In women, hair loss begins after
menopause in particular.
[0008] Depending on the root cause of baldness, hair can grow back
more or less easily. There is no cure-all treatment and each case
of hair loss is complex and has to be treated appropriately.
[0009] In the case of androgenetic loss, the main cause of baldness
in both men and women, rapid and effective treatment is necessary.
If nothing is done, gradual atrophy of the hair follicles and roots
takes place as well as a decrease in hair diameter and alopecia
develops. There is no curative treatment for androgenetic alopecia.
It actually results from the hair follicle being over-sensitive to
dihydrotestosterone (DHT) which shortens the capillary growth
phase. Administration of oral antiandrogens such as Finasteride
(Propecia.RTM.), which blocks the enzyme responsible for
transforming testosterone into dihydrotestosterone in the hair
follicles, reduces hair loss and activates re-growth. However,
while many hopes were pinned on this product, it is contraindicated
in women and recent warnings have been issued with regard to its
use in men. In fact, the results of a clinical study suggest that
there is a risk of developing a serious form of cancer of the
prostate in patients taking this drug.
[0010] Minoxidil (Rogaine.RTM.), another anti-hair loss preparation
currently available on the market, is only useful at the start of
alopecia. Its application to the scalp delays the end of the hair
growth cycle, and therefore hair loss, but it does not reconstruct
inactive follicles. Hair loss recommences a few weeks after
stopping treatment.
[0011] There are also cosmetic products containing agents which
confer volume on thinning hair. However, these agents do not
stimulate growth. They can nonetheless give the impression of
thicker hair since they coat the hair shaft and thus increase its
diameter. These products only offer a temporary solution since they
disappear each time hair is washed.
[0012] It would therefore be useful to provide the cosmetic or
pharmaceutical industry with an effective product that is devoid of
side effects to stimulate hair growth and/or prevent hair loss.
Such a product would therefore reduce, and possibly even prevent,
alopecia.
[0013] The Applicant has discovered a natural tetrapeptide and
analogues that stimulate vital hair functions, in particular by
activating epithelial stem cells, and therefore to achieve the
desired objective. These derivatives also have the advantage of
being obtained by peptide synthesis routes that are easy to
implement and which are therefore not costly. Moreover, these
compounds also have very low, and possibly even no, toxicity for
the body.
[0014] The peptides or analogues used within the scope of this
invention are derivatives with the basic structure
Acetyl-Ser-Asp-Lys-Pro (AcSDKP). Their therapeutic properties are
well-established (WO-88/00594 and WO-97/28183).
[0015] The Applicant has recently demonstrated their angiogenic
properties (WO-02/24218).
[0016] No documents of the prior art describe or suggest that these
compounds could have a cosmetic anti-hair loss effect nor that
their use could lead to a positive effect in stimulating stem cells
present in the hair follicles.
[0017] This invention therefore concerns the use of at least one
compound of formula (I):
##STR00001##
in which
[0018] A.sub.1 is the radical corresponding to D- or L-Ser,
[0019] A.sub.2 is the radical corresponding to D- or L-Asp or
Glu,
[0020] A.sub.3 is the radical corresponding to D- or L-Lys, Arg or
Orn,
[0021] A.sub.4 is the radical corresponding to D- or L-Pro,
[0022] R.sub.1 and R.sub.2 are independently chosen from among the
hydrogen atom, linear or branched substituted or non-substituted
C.sub.1-C.sub.12 alkyl groups, substituted or non-substituted
linear or branched substituted or non-substituted C.sub.7-C.sub.20
arylalkyl groups, R.sub.4CO-- and R.sub.4COO-- where R.sub.4 is a
linear or branched substituted or non-substituted C.sub.1-C.sub.12
alkyl group, or a substituted or non-substituted C.sub.7-C.sub.20
arylalkyl, where substitutions include OH, NH.sub.2 and COOH,
[0023] X.sub.1 and X.sub.2 are peptide or pseudopeptide bonds,
[0024] X.sub.3 is a radical chosen from among --CO and
--CH.sub.2--, and
[0025] R.sub.3 is a group chosen from among --OH, --NH.sub.2,
linear or branched alcoxy C.sub.1-C.sub.12 or
--NH--X.sub.4--CH.sub.2-Z, where X.sub.4 is a linear or branched
C.sub.1-C.sub.12 hydrocarbon group and Z is a hydrogen atom or
--OH, --CO.sub.2H or --CONH.sub.2, as well as their physiologically
acceptable salts, in a composition as an agent for controlling hair
loss.
[0026] Among the alkyl groups particularly suited to implementation
of this invention, linear or branched C.sub.1-C.sub.6 alkyl groups
are preferred. In particular, the methyl, ethyl, propyl, isopropyl,
butyl, isobutyl and tertbutyl groups can be cited.
[0027] The term aryl group in the context of this invention relates
to an aromatic carbon group with 6 to 14 carbon atoms. The phenyl,
naphthyl and anthracenyl groups can be cited as examples of
this.
[0028] The preferred arylalkyl groups of the invention include the
benzyl and phenethyl groups.
[0029] The peptides or pseudopeptides corresponding to formula (I)
are derived form the basic tetrapeptide structure
Acetyl-Ser-Asp-Lys-Pro (AcSDKP).
[0030] The term "radical corresponding to" refers to the radical A
of formula:
[0031] NH.sub.2--CH(A)-COOH corresponding to the amino acid.
[0032] Thus, A is:
[0033] --CH.sub.2OH for Ser,
[0034] --CH.sub.2COOH for Asp,
[0035] --CH.sub.2--CH.sub.2--COOH for Glu,
[0036] --(CH.sub.2).sub.3--NH--C(NH)NH.sub.2 for Arg,
[0037] --(CH.sub.2).sub.3--NH.sub.2 for Orn and
[0038] --(CH.sub.2).sub.4--NH.sub.2 for Lys
[0039] for the terminal amino acid A.sub.4, this consists of the
structure:
[0040] .dbd.N--CH(A)-CO-- or NH--(CH)A-CO--.
[0041] The term "pseudopeptide" refers to compounds that are
similar to the reference peptide but in which one or more peptide
--CO--NH-- bonds have been replaced by a bond equivalent to the
peptide bond called pseudopeptidic, that is --CH.sub.2--NH--,
--CH.sub.2--S--, --CH.sub.2--O--, --CO--CH.sub.2--,
--CH.sub.2--CO--, --CH.sub.2--CH.sub.2-- represented by
.psi.(CH.sub.2NH) for example.
[0042] Among the R.sub.1 and R.sub.2 radicals, the hydrogen atom or
R.sub.4CO-- radicals are particularly preferred where R.sub.4
represents a C.sub.1-C.sub.3 alkyl group, namely CH.sub.3CO as well
as HOOC--CH.sub.2--CH.sub.2--CO--O,
Similarly, R.sub.3 is preferably NH.sub.2, OH or NHCH.sub.3.
[0043] Among the compounds of formula (I) suited to implementation
of the invention, the following can be cited:
[0044] CH.sub.3CO-Ser-Asp-Lys-Pro-OH,
[0045] CH.sub.3CO-Ser-.psi.-(CH.sub.2NH)-Asp-Lys-Pro-OH
[0046] CH.sub.3CO-Ser-Asp-.psi.-(CH.sub.2NH)-Lys-Pro-OH
[0047] CH.sub.3CO-Ser-Asp-Lys-.psi.-(CH.sub.2N)-Pro-OH
[0048] CH.sub.3CO-Ser-.psi.-(CH.sub.2NH)-Asp-Lys-Pro-NH.sub.2
[0049] CH.sub.3CO-Ser-Asp-.psi.-(CH.sub.2NH)-Lys-Pro-NH.sub.2
[0050] CH.sub.3CO-Ser-Asp-Lys-.psi.-(CH.sub.2N)-Pro-NH.sub.2
[0051] H-Ser-.psi.-(CH.sub.2NH)-Asp-Lys-Pro-OH
[0052] H-Ser-Asp-.psi.-(CH.sub.2NH)-Lys-Pro-OH
[0053] H-Ser-Asp-Lys-.psi.-(CH.sub.2N)-Pro-OH
[0054]
HOOCCH.sub.2CH.sub.2CO-Ser-.psi.-(CH.sub.2NH)-Asp-Lys-Pro-OH
[0055]
HOOCCH.sub.2CH.sub.2CO-Ser-Asp-.psi.-(CH.sub.2NH)-Lys-Pro-OH
[0056]
HOOCCH.sub.2CH.sub.2CO-Ser-Asp-Lys-.psi.-(CH.sub.2NH)-Pro-OH
[0057] H-Ser-.psi.-(CH.sub.2NH)-Asp-Lys-Pro-NH.sub.2
[0058] H-Ser-Asp-.psi.-(CH.sub.2NH)-Lys-Pro-NH.sub.2
[0059] H-Ser-Asp-Lys-.psi.-(CH.sub.2N)-Pro-NH.sub.2
[0060]
HOOCCH.sub.2CH.sub.2CO-Ser-.psi.-(CH.sub.2NH)-Asp-Lys-Pro-NH.sub.2
[0061]
HOOCCH.sub.2CH.sub.2CO-Ser-Asp-.psi.-(CH.sub.2NH)-Lys-Pro-NH.sub.2
[0062]
HOOCCH.sub.2CH.sub.2CO-Ser-Asp-Lys-.psi.-(CH.sub.2N)-Pro-NH.sub.2
[0063] CH.sub.3CO-Ser-Asp-Lys-Pro-NH.sub.2
[0064] H-Ser-Asp-Lys-Pro-NH.sub.2
[0065] CH.sub.3CO-Ser-Asp-Lys-Pro-NHCH.sub.3
[0066] H-Ser-Asp-Lys-Pro-NHCH.sub.3
[0067] HOOCCH.sub.2CH.sub.2CO-Ser-Asp-Lys-Pro-NHCH.sub.3
[0068] HOOCCH.sub.2CH.sub.2CO-Ser-Asp-Lys-Pro-NH.sub.2
[0069] A compound of formula (I) particularly suited to
implementation of this invention is the natural tetrapeptide
CH.sub.3CO-Ser-Asp-Lys-Pro (AcSDKP).
[0070] The term "physiologically acceptable salt" in the context of
this invention refers to any salt prepared from any physiologically
acceptable non-toxic acid, including organic and inorganic acids.
Such acids include acetic, benzenesulphonic, benzoic, citric,
ethanesulphonic, fumaric, gluconic, glutamic, hydrobromic,
hydrochloric, lactic, maleic, malic, mandelic, methanesulphonic,
mucic, nitric, pamoic, pantothenic, phosphoric, succinic, tartaric
and paratoluenesulphonic acids. Advantageously, hydrochloric acid
is used.
[0071] The compounds of formula (I) defined above have been shown
to have a stimulating effect on cell growth and restructuring the
epithelial sheath of hair follicles which constitute the source of
stem cells. A large increase in the number of neo-keratinocytes and
undifferentiated cells in the pilar structures of hair treated with
compounds of formula (I) has been observed.
[0072] The Applicant has found that at the molecular level the
effects of compounds used in accordance with the invention results
in increased collagen IV and laminin 5 expression along the length
of the hair, with a more pronounced effect in the median section
situated between the bulge and the bulb.
[0073] One embodiment of the invention concerns the cosmetic use of
a compound of formula (I) as defined earlier in a composition as an
agent to trigger the growth and/or differentiation of stem cells
and/or neo-keratinocytes in the epithelial sheath of hair and/or to
stimulate the production of collagen IV and/or laminin 5 in the
hair follicle.
[0074] More particularly, this invention concerns the cosmetic use
of a compound of formula (I) as defined earlier in a composition as
an agent to trigger and/or stimulate hair growth.
[0075] In another aspect of the invention, the compounds of formula
(I) as defined above can be used to increase hair density.
[0076] Another particular aspect of the invention concerns the use
of compounds of formula (I) as defined above in the prevention of
alopecia.
[0077] The natural tetrapeptide CH.sub.3CO-Ser-Asp-Lys-Pro (AcSDKP)
(WO-88/00594) can be obtained by means of conventional peptide
synthesis. The peptides or pseudopeptides of formula (I) related to
the AcSDKP derivative can also be obtained by means of peptide or
pseudopeptide synthesis as described in the document
WO-97/28183.
[0078] The compounds of formula (I) are present in the compositions
used to implement this invention in amounts ranging from 0.001% to
10% by weight, preferably between 0.005% and 5% by weight with
respect to the total weight of the composition.
[0079] The cosmetic compositions used according to the invention
are intended for topical use and can be in any pharmaceutical form
conventionally used for this type of application, and namely in the
form of emulsions (oil-in-water, water-in-oil, oil-in-water-in-oil
or water-in-oil-in-water triple emulsions), aqueous gels, aqueous
solutions, hydroalcoholic or oily solutions. They can be more or
less fluid and in the form of a white or coloured cream, ointment,
milk, lotion, serum, paste, mousse or biphase. They can also be in
the form of an aerosol. The compositions used to implement this
invention can be in the form of a lotion, gel, soap, aerosol,
shampoo or mousse.
[0080] The compositions used within the scope of this invention
contain derivatives contain, in addition to derivatives of formula
(I), one or more excipients which can be chosen from among
compounds with good compatibility with the active ingredients
present in the formula. This can be, for example, natural polymers
such as polysaccharides (xanthan gum, carob gum, peptin, etc.) or
polypeptides, cellulose derivatives such as methylcellulose,
hydroxypropylcellulose, hydroxypropylmethylcellulose, or synthetic
polymers, polaxamers, carbomers, PVA or PVP.
[0081] Finally, the compositions of the invention can also contain
various other excipients of the co-solvent type such as ethanol,
glycerol, benzyl alcohol, wetting agents (glycerol), agents to
facilitate diffusion (transcurol, urea) or even antibacterial
preservatives (0.15% methyl p-hydroxybenzoate). They can also
contain surfactants, stabilisers, emulsifiers, thickeners, other
active ingredients generating a complementary effect or possibly
synergetic effect, trace elements, essential oils, fragrances,
colouring agents, collagen, chemical or mineral filters, hydrating
agents or thermal spa waters.
[0082] In a particular embodiment of the invention, the derivatives
of formula (I) are combined with at least one other active
ingredient.
[0083] This invention also concerns a cosmetic process for the
treatment and/or prevention of hair loss, particularly alopecia,
consisting of the application to the scalp of a composition
containing at least one derivative of formula (I) as defined
earlier.
[0084] The following examples are given solely for the purpose of
illustrating the invention and in no way limit it.
EXAMPLE 1
Experimental Study
[0085] Study of the stimulating activity of tetrapeptide AcSDKP on
the growth of isolated hairs maintained in a state of ex vivo
survival. This study was conducted on 66 human hair explants
obtained from scalp-plasty.
Method
[0086] Hairs comprised of hair follicles were isolated by micro
dissection and individually placed in wells (96-well culture plate)
and maintained in survival for 15 days under conventional tissue
culturing conditions (37.degree. C., 5% CO.sub.2) in Williams
medium.
[0087] Tetrapeptide AcSDKP added to the culture medium was tested
at three concentrations: 10.sup.-4 M, 10.sup.-7 M and 10.sup.-10 M.
Minoxidil.RTM. (2,4-diamino 6-piperidinopyrimidine 3-oxide) at a
concentration of 10.sup.-5 M was used as a positive control.
[0088] Culture mediums containing the compounds to be tested were
renewed every three days.
[0089] Hairs were photographed using a microscope and a CCD camera
coupled to acquisition software and archiving.
[0090] Each hair was measured on D0, D3, D6, D8, D10, D13 and D15
using a LEICA IM1000 measurement module to give measurements in
.mu.m.
[0091] On D8 and D15 of treatment, six hairs from each batch
(untreated control, Minoxidil.RTM., +AcSDKP 10.sup.-4 M, +AcSDKP
10.sup.-7 M, +AcSDKP 10.sup.-10 M) were removed and prepared for
histological study.
[0092] Three explants from each batch were fixed in ordinary Bouin
and the three other hairs were frozen and stored at -80.degree.
C.
[0093] In addition to measurements of the length of each hair, the
general morphology of each hair was analysed and the expression of
laminin 5 and collagen IV, two constituents of the basal membrane,
was evaluated. Moreover, immunolabeling with anti-Ki67 antibodies
(nuclear protein expressed in growing cells) was carried out of
cells in mitosis in order to establish the mitotic index of
follicle structures.
Results
1. Size of Hairs
[0094] Each hair was photographed and its length evaluated using
software giving values in .mu.m. Observations made on D15 of
treatment for the set of explants tested shows that the presence of
AcSDKP at 10.sup.-10 M in the culture medium significantly
stimulates hair growth (FIG. 1). On the other hand, no significant
difference in the hair shafts was found in the presence of AcSDKP
tested at higher concentrations.
2. Cell Proliferation
[0095] This study was conducted on cuts of frozen hairs.
Immunolabeling of the cells present along the epithelial sheath
with anti-Ki67 antibodies reveals a mitogenic effect of AcSDKP. The
results presented in Table 1 show that AcSDKP at the three
concentrations tested significantly increases the number of cells
in mitosis in treated hair compared to control hair. Measurements
on D8 of treatment show that this effect is maximal for AcSDKP at
10.sup.-10 M (+300%). Evaluation of the number of cells in mitosis
also shows that the stimulating effect of AcSDKP is significantly
greater than that found for Minoxidil.RTM. at 10.sup.-5 M (+38%),
especially at D8 of treatment.
[0096] The observations made with these hairs therefore demonstrate
that AcSDKP ex vivo triggers cell proliferation more rapidly than
Minoxidil.RTM. in the external epithelial sheaths, regions where
stem cells are located when hair is in the growth phase.
TABLE-US-00001 TABLE 1 Average number of cells in mitosis per mm of
hair Minoxidil .RTM. AcSDKP AcSDKP AcSDKP Days Control 10.sup.-5 M
10.sup.-4 M 10.sup.-7 M 10.sup.-10 M D0 12.6 D8 13.4 18.5 35.4 34
52.6 D15 24.5 23.3 23.4 21.5 29.3
3. General Morphology
[0097] Analysis of the general morphology of histological cuts
stained with Masson Trichromium focusing on the area between the
opening of the sebaceous glands and the hair bulb.
[0098] Hairs cultured on D0 naturally undergo changes during
survival as is observed after an in vivo graft. The restructuring
effect of AcSDKP was visualised through the appearance of
neo-keratinocytes along the conjunctival sheath which migrate from
the bulge and to a lesser extent from the bulb, two regions where
stem cells are found. Observation of the set of isolated hairs
shows that the presence of AcSDKP in the culture medium leads to
restructuring of the epithelial sheaths as of D8 of treatment. This
effect leads to clear appearance of neo-keratinocytes along the
length of the hair, from the epidermis to the bulb. The structure
of the neo-keratinocytes formed in this way is uniform and
stratified. It varies as a function of treatment time and AcSDKP
concentration. The clearest activity was found on D15 of treatment
with AcSDKP at 10.sup.-10 M.
[0099] On D8, 2 to 3 neo-keratinocyte sites were visualised in the
control hair but were absent from explants treated with
Minoxidil.RTM. at 10 .mu.M as well as with AcSDKP 10.sup.-4 M. On
the other hand, there was a very clear appearance of
neo-keratinocytes (5 to 6 cell bases) throughout the length of
hairs treated with AcSDKP at 10.sup.-7 M and 10.sup.-10 M. It
should be pointed out that this epithelium adheres fully to the
underlying collagen in hairs kept in survival in the presence of
AcSDKP tested at the two active concentrations whereas the
epithelial sheaths are more or less detached from collagen in the
controls as well as in the presence of AcSDKP at 10.sup.-4 M and
Minoxidil.RTM. (FIG. 2A).
[0100] On D15, neo-keratinocytes form a uniform and moderately
stratified structure in treated hairs. This is slightly more
structured with Minoxidil.RTM. at 10.sup.-5 M compared to untreated
hair. This becomes increasingly thicker with AcSDKP at 10.sup.-4 M,
10.sup.-7 M and especially with 10.sup.-10 M, with 6 to 7 cell
bases close to the bulb, with up to 11 to 12 in the bulge (FIG.
2B).
4. Expression of Basal Membrane Collagen (Type IV)
[0101] Immunolabeling of type IV collagen was carried out on frozen
cuts of the control hairs and hairs treated with AcSDKP or
Minoxidil.RTM. in order to quantify this constituent of the basal
membrane which separates the dermal compartment from the epithelial
compartment. Analysis of the results obtained revealed a minimally
stimulating effect of AcSDKP on expression of this protein in hair
kept in survival in the presence of the tetrapeptide. In fact, as
of D8 of treatment with AcSDKP at the three concentrations tested,
there is a very clear increase in collagen IV in the bulb, and
especially in the median section of hair at a point between the
bulge and bulb. The increase in collagen levels found on D8 and D15
is all the more marked for AcSDKP tested at a concentration of
10.sup.-10 M. This expression of collagen IV is often less evident
in hair treated with Minoxidil.RTM. (FIGS. 3A and 3B).
5. Expression of Laminin 5
[0102] Laminin 5, another constituent of the basal membrane, is
present in the bulb around the dermal papilla as well as in the
median section of hair. Immunolabelling of laminin 5 revealed a
significant increase in the level of this protein in hair
structures following exposure of hair to AcSDKP. This effect was
considerably more marked in the median zone of hair treated with
AcSDKP at a concentration of 10.sup.-10 M (FIG. 4A) on D8 of
treatment. Nevertheless, expression of laminin 5 was higher on D15
with AcSDKP at 10.sup.-7 M (FIG. 4B).
6. Expression of Keratin 19 (CK19), an Epithelial Stem Cell
Marker
[0103] Immunolabeling of keratin 19 (CK19) was carried out on
frozen cuts of control hairs and hairs treated with AcSDKP or
Minoxidil.RTM. in order to evaluate the proportion of
undifferentiated primitive cells (or even stem cells) present in
the external epithelial sheaths of the hair follicles. The results
obtained show that expression of CK19 is significantly greater in
hair kept in survival in the presence of the tetrapeptide compared
to control hair and especially with respect to hair treated with
Minoxidil.RTM.. The increase in CK19 levels on D8 and D15 of
treatment is maximal for AcSDKP tested at a concentration of
10.sup.-10 M (FIG. 5). Analysis of the results obtained shows a
stimulating effect of AcSDKP in epithelial stem cells distributed
along the external sheath of the follicle and responsible for
renewal of the hair cycle.
7. Tissue Localisation of Radio Labelled AcSDKP in Animals Having
Received a Single Dose In Vivo of the Tetrapeptide
[0104] A complementary study using in vivo labelling of target
organs of the peptide was carried out. This study was developed
using [.sup.3H]AcSDKP (100 Ci/mmole) specifically tritiated in the
lysine lateral chain. In vivo injection of rats with
[.sup.3H]AcSDKP followed by autoradiography of cuts taken from the
whole animal made it possible to visualise accumulation of the
molecule in the animal's skin as well as in the hair follicles of
whiskers (FIG. 6). This preferential site for AcSDKP suggests that
the biological effect of this tetrapeptide is probably exerted in
these areas.
EXAMPLE 2
Lotion
TABLE-US-00002 [0105] Ethyl alcohol 50% AcSDKP 0.1% Water 49.9%
Fragrance q.s.
EXAMPLE 3
Lotion
TABLE-US-00003 [0106] Hydroxyethylcellulose 0.4% Ethyl alcohol 25%
Butane 1,3-diol 38.4% Paramethylbenzoate 0.2% AcSDKP 0.05%
Fragrance 1% Water q.s. for 100%
EXAMPLE 4
Oil-in-Water Emulsion
Oily Phase
TABLE-US-00004 [0107] Vitamin E 1% Sorbitan monoleate 2%
Quaternum-18 0.5% Paraffin 6.5%
Aqueous Phase
TABLE-US-00005 [0108] AcSDKP 0.5% Xanthan gum 1% Preservative 0.3%
Fragrance q.s. Water q.s. for 100%
* * * * *